ciclo: XXXIII

tutor: Prof. Rita Canipari
relatore: Antonio Capalbo

Titolo della tesi: Non-invasive analysis of the embryonic genome for the development of improved preimplantation genetic testing (PGT) protocols

To characterize cell-free DNA origin and determine whether spent blastocyst medium (SBM) is a suitable template for non-invasive genomic assessment of IV§F–generated embryos. As preliminary phase, the origin of aneuploidy was investigated in human blastocysts by using NGS analysis on multiple TE and ICM biopsies from the same embryo. We concluded that whole chromosome aneuploidies are mostly meiotic in origin and can be targeted from SBM samples. On the contrary, we provide definitive evidence about the preferential mitotic nature of segmental aneuploidies, making them as an unsuitable target for SBM analysis. Next, we have assessed the concordance rate and genotyping accuracy of SBM vs embryonic biopsies under standard genetic and IVF laboratory protocols. Under this setting we have highlighted poor predictive performance of embryonic culture media in defining the genetic and chromosomal constitution of the embryo. In particular, In PGT-M tests, for BF and SBM, 2.9% and 20.8% of all samples, respectively, produced a diagnosis concordant with the corresponding TE (n=2 of 69 and 15 of 72, respectively). In PGT-A tests, BF analysis showed high amplification failure rates (65.2%) and an overall concordance rate of 37.5% among amplified samples. After improving embryo culture conditions and genetic protocols for the analysis of SBM samples, we could obtained concordant chromosome analysis between SBM and their relative TE biopsies as high as 87% (N=115) from day 6 embryos in a prospective pilot multicenter study. With the new optimized strategy, a prospective multicenter trial on 1301 paired TE and SBM sample was performed. Results have confirmed high concordance and reproducibility among the IVF centers involved. Improvement in embryo culture and genetic technologies have shown the possibility to utilize SBM for non-invasive assessment of embryonic genetics by the analysis of cfDNA. Future clinical trials are warranted to assess clinical utility of the novel strategy that holds the potentiality of becoming a milestone achievement in the field of preimplantation genetics.

Produzione scientifica

  • 11573/1118428 - 2017 - Human female meiosis revised: New insights into the mechanisms of chromosome segregation and aneuploidies from advanced genomics and time-lapse imaging (01a Articolo in rivista)
  • 11573/1118442 - 2017 - Prevention of neural tube defects and maternal gestational diabetes through the inositol supplementation: preliminary results (01a Articolo in rivista)
  • 11573/1183954 - 2018 - Erratum to: Segregation of mitochondrial DNA heteroplasmy through a developmental genetic bottleneck in human embryos (Nature Cell Biology, (2018), 20, 2, (144-151), 10.1038/s41556-017-0017-8) (01a Articolo in rivista)
  • 11573/1173223 - 2018 - Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions (01a Articolo in rivista)
  • 11573/1173220 - 2018 - Biochemical pregnancy loss after frozen embryo transfer seems independent of embryo developmental stage and chromosomal status (01a Articolo in rivista)
  • 11573/1311293 - 2019 - Definition and validation of a custom protocol to detect miRNAs in the spent media after blastocyst culture: searching for biomarkers of implantation (01a Articolo in rivista)
  • 11573/1015148 - 2017 - Abnormally fertilized oocytes can result in healthy live births: improved genetic technologies for preimplantation genetic testing can be used to rescue viable embryos in in vitro fertilization cycles (01a Articolo in rivista)
  • 11573/932326 - 2016 - Human Embryos Created by Embryo Splitting Secrete Significantly Lower Levels of miRNA-30c (01a Articolo in rivista)
  • 11573/434020 - 2012 - The 312N variant of the luteinizing hormone/choriogonadotropin receptor gene (LHCGR) confers up to 2.7-fold increased risk of polycystic ovary syndrome in a Sardinian population (01a Articolo in rivista)
  • 11573/1337528 - 2019 - Embryonic cell-free DNA versus trophectoderm biopsy for aneuploidy testing: concordance rate and clinical implications (01a Articolo in rivista)
  • 11573/932328 - 2016 - Pre-implantation genetic testing in ART: who will benefit and what is the evidence? (01a Articolo in rivista)
  • 11573/853041 - 2014 - Secreted miRNAs can be profiled with high accuracy and reproducibility from blastocyst spent culture media: a new potential biomarker for non-invasive embryo selection (04d Abstract in atti di convegno)
  • 11573/1015149 - 2017 - Preimplantation genetic diagnosis for aneuploidy testing in women older than 44 years: a multicenter experience (01a Articolo in rivista)
  • 11573/853032 - 2016 - Consistent and reproducible outcomes of blastocyst biopsy and aneuploidy screening across different biopsy practitioners: a multicentre study involving 2586 embryo biopsies (01a Articolo in rivista)
  • 11573/853046 - 2015 - Blastocyst morphology does not significantly increase the selection between euploid blastocysts in frozen embryo transfer cycles: evidences from a prospective non-selection study (04d Abstract in atti di convegno)
  • 11573/902743 - 2016 - Electronic witness system in IVF—patients perspective (01a Articolo in rivista)
  • 11573/614648 - 2015 - No evidence of association between blastocyst aneuploidy and morphokinetic assessment in a selected population of poor-prognosis patients: a longitudinal cohort study (01a Articolo in rivista)
  • 11573/1246631 - 2019 - An integrated investigation of oocyte developmental competence: expression of key genes in human cumulus cells, morphokinetics of early divisions, blastulation, and euploidy (01a Articolo in rivista)
  • 11573/872506 - 2016 - Follicular versus luteal phase ovarian stimulation during the same menstrual cycle (DuoStim) in a reduced ovarian reserve population results in a similar euploid blastocyst formation rate: new insight in ovarian reserve exploitation (01a Articolo in rivista)
  • 11573/1383455 - 2020 - Incidence, origin, and predictive model for the detection and Clinical Management of segmental aneuploidies in human embryos (01a Articolo in rivista)
  • 11573/853078 - 2015 - Reduction of multiple pregnancy rate in advanced maternal age population after the introduction of an elective single embryo transfer policy: a pre- and post- intervention study (04d Abstract in atti di convegno)
  • 11573/1337514 - 2019 - Chromosome errors in human eggs shape natural fertility over reproductive life span (01a Articolo in rivista)
  • 11573/853036 - 2016 - The impact of biopsy on human embryo developmental potential during preimplantation genetic diagnosis (01g Articolo di rassegna (Review))
  • 11573/1337526 - 2019 - Optimizing clinical exome design and parallel gene-testing for recessive genetic conditions in preconception carrier screening: Translational research genomic data from 14,125 exomes (01a Articolo in rivista)
  • 11573/1189480 - 2018 - Prevalence of XXY karyotypes in human blastocysts: multicentre data from 7549 trophectoderm biopsies obtained during preimplantation genetic testing cycles in IVF (01a Articolo in rivista)
  • 11573/1341044 - 2019 - Time of morulation and trophectoderm quality are predictors of a live birth after euploid blastocyst transfer: a multicenter study (01a Articolo in rivista)

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